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Examination of ethylacetate extracts of media fermented by Streptomyces antibioticus 3720 has shown the presence of a number of acidic substances related to actinomycin. One of these has been isolated in pure form and found to be a mono-lactone of actinomycin. The fermented medium (a modification of that of Goss and KatzJ) was adjusted to pH 7.0 and extracted with 1/3 volume of ethylacetate. The aqueous phase was then adjusted with H2SO4 to pH 2.5 and extracted twice with 1/3 volumes of ethylacetate. These latter extracts were pooled, concentrated to a small volume, dried over anhydrous Na2SO4, and concentrated to 100 ml. This solution was then extracted with 0.1 M pH 7.0 phosphate buffer (to remove the acidic components), and the buffer solution was back extracted with ethylacetate. The mono-lactone in this last extract was separated from residual actinomycin and other fermentation products by column chromatography with Sephadex LH-20 as support and methanol as eluant. The purity of the middle fraction, which contained the mono-

ACTINOMYCIN MONO-LACTONE, A METABOLITE OF STREPTOMYCES ANTIBIOTICUS 3720